CN1214633A - Stabilised growth hormone formulation and method of preparation thereof - Google Patents
Stabilised growth hormone formulation and method of preparation thereof Download PDFInfo
- Publication number
- CN1214633A CN1214633A CN97193316A CN97193316A CN1214633A CN 1214633 A CN1214633 A CN 1214633A CN 97193316 A CN97193316 A CN 97193316A CN 97193316 A CN97193316 A CN 97193316A CN 1214633 A CN1214633 A CN 1214633A
- Authority
- CN
- China
- Prior art keywords
- preparation
- hgh
- growth hormone
- buffer
- stabilizing agent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/27—Growth hormone [GH], i.e. somatotropin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/38—Cellulose; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/06—Drugs for disorders of the endocrine system of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Endocrinology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Inorganic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dermatology (AREA)
- Organic Chemistry (AREA)
- Diabetes (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
A method for the preparation of a stable, liquid formulation of growth hormone, comprising growth hormone, a buffer and a stabilising effective amount of at least one stabilising agent selected from the group consisting of: (i) polyethylene-polypropylene glycol non-ionic surfactants, (ii) taurocholic acid or salts or derivatives thereof, and (iii) methyl cellulose derivatives, wherein the method comprises admixing the growth hormone with the buffer and the stabilising agent(s) under conditions such that the growth hormone is not exposed to concentrations of the buffer or stabilising agent(s) which are greater than 2x the final concentrations of the buffer or stabilising agent(s) in the formulation.
Description
Invention field
The present invention relates to growth hormone (GH) preparation of stabilisation, especially stabilized human growth hormone (hGH) liquid preparation by mixing the stabilisation excipient.The liquid preparation of these hGH has the chemistry and the physical stability of improvement.The invention particularly relates to the method for these stable GH preparations of preparation.
The background technology of invention
Comprise about 191 the amino acid whose protein of the growth hormone of humans and animals in antepituitary, finding.The main biological action of GH is to promote sign to form in childhood in the humans and animals, keeps tissue in adult organism.The organ that influenced by GH comprises skeleton, muscle, connective tissue and internal organs.Growth hormone by with target cell membrane on specific receptor interact and to work.
Human growth hormone (hGH) is a kind of critical hormone that participates in regulating normal human's growth, and influences various physiology and metabolic function, and comprising linear osteogenesis, lactogenic and cellular energy use.The shortage of child hGH causes of short and small stature, and this disease is used hGH by external source and obtained medical treatment.
In the past, make great efforts to concentrate on the molecular function of the growth hormone of determining each kind.Commercial interest from medical science and veterinary applications is very strong, and has cloned the hGH gene.Just in mammal and bacterial cell culture system, prepare hGH and its derivant methionyl-hGH (met-hGH) at present with biosynthesis.
For medicine for treatment thing preparation at commercial supply hGH, must the preparation stable formulation.These preparations must keep active at suitable duration of storage; They must be easy to preparation, and can be taken like a shot by patient and use.
Industry has been prepared people GH in every way.For example, U.S. Patent number 5,096,885 disclose except hGH, the acceptable hGH preparation of stable materia medica that also comprises glycine, mannitol, buffer and optional a kind of non-ionic surface active agent, wherein hGH: the mol ratio of glycine is 1: 50-200.International Patent Publication No. WO 93/19776 discloses the GH preparation that injectable is used, it comprises citrate as buffer substance, with optional somatomedin such as insulin like growth factor or epidermal growth factor, aminoacid such as glycine or alanine, mannitol or other sugar alcohol, glycerol and/or antiseptic such as benzyl alcohol.International Patent Publication No. WO 94/01398 discloses and has comprised hGH, buffer, the GH preparation of non-ionic surface active agent and mannitol, neutral salt and/or the antiseptic chosen wantonly.
In European Patent Publication No 0131864 (with corresponding Australian Patent numbers 579016); a kind of molecular weight is disclosed greater than 8500 daltonian proteic aqueous solutions; wherein make these albumen protected, do not prevent degeneration and albumen precipitation thereby be not adsorbed at the interface by the surfactant that comprises linear polyoxyalkylene chain that adds as stabilizing agent.
European Patent Publication No 0211601 discloses a kind of preparation of growing of promoting, its comprise growth promoting hormones and contain polyoxyethylene-polyoxypropylene unit, mean molecule quantity is about 1,100-about 40, the aqueous mixture of 000 block copolymer, wherein said preparation has kept the mobile and biological activity when using of growth promoting hormones.European Patent Publication No 0303746 subsequently discloses various other stabilizing agents of growth promoting hormones in aqueous environment, comprises some polyhydric alcohol, aminoacid, has the amino acid polymer and the choline salt of charged side group under physiological pH.
The pharmaceutical preparation of hGH is generally unstable, especially in solution.Physical instability may cause producing the chemical degradation thing, and as the hGH of deamidate or sulfoxylate form, dimer or high molecular are assembled thing ((1988) biotechnology such as Becker and applied biochemistry 10,326; Pearlman and Nguygen (1989), at D.Marshak and D.Liu (volume), therapeutic peptide and protein, preparation, send with target fixed, modern molecular biology communication, cold spring harbor laboratory, cold spring port, New York, pp23-30; (1987) biotechnology such as Becker and applied biochemistry 9,478).
Because hGH is unstable in solution, the pharmaceutical preparation of hGH is tending towards existing with the freeze-dried that must prepare again before use.Lyophilization is used in some wherein biological activity and biochemistry integrity to keep polypeptide under the stable not enough holding conditions in solution usually, yet it will be favourable avoiding lyophilization, because it is a costliness and production stage consuming time.Usually before use by adding the materia medica acceptable diluent, prepare the freeze-dried preparation of hGH again as acceptable diluent on Injectable sterile water, physiological saline solution or the suitable sterile physiological.Preferably the obtain solution again of hGH is housed under 4 ℃ so that chemistry and mechanical degradation reaction are reduced to minimum, yet, some degradeds will be produced the longest reaching in shelf-life of 14 days.
With the hGH pharmaceutical preparation that liquid form provides, it will be particularly favourable especially keeping the pharmaceutical preparation of hGH stability in long-time.As mentioned above, present liquid preparation is in the chemistry that is subjected on the storage time to produce in processing and storage and the restriction of mechanical degradation product.In (1987) (the same) such as Becker, reported with dimer to form relevant problem, and the not success of effort of avoiding the hGH dimer to form in the past.
An object of the present invention is to provide and do not cause forming undesirable aggregation or do not cause reducing biological activity or the stable hGH liquid preparation of the chemical change of change receptor identification.Another purpose provides and can send by means of being used for hypodermic needleless injector, maybe can be made into to be used for the preparation of the aerosol of pulmonary.
Summary of the invention
According to the present invention; a kind of method for preparing stable growth hormone liquid preparation is provided; wherein said preparation comprises growth hormone; buffer and stabilisation effective dose at least a is selected from following stabilizing agent: (ⅰ) polyethylene glycol-propylene glycol non-ionic surface active agent; (ⅱ) taurocholic acid or its salt or derivant; (ⅲ) methylcellulose derivant; wherein this method is included in growth hormone and is not exposed under the condition greater than the buffer of buffer in 2 * preparation or stabilizing agent ultimate density or stabilizer concentration, and growth hormone is mixed with buffer and stabilizing agent.
The invention still further relates to stable growth hormone liquid preparation by the method preparation of summarizing above.
Another aspect the invention still further relates to a kind of stable growth hormone liquid preparation, and said preparation comprises at least a stabilizing agent that is selected from taurocholic acid or its salt or derivant and methylcellulose derivant of growth hormone, buffer and stabilisation effective dose.
Unless this paper has explanation in addition, in this manual, word " comprises " or its version was interpreted as referring to comprise described key element or key element group as " comprising ", but does not get rid of any other key element or key element group.
In an especially preferred example, the invention provides a kind of method for preparing the stable acceptable human growth hormone's of materia medica liquid preparation, said preparation comprises: (a) hGH of materia medica effective dose, (b) 0.01-5.0%w/v at least a is selected from stabilizing agent and (c) the acceptable buffer of materia medica in the top general introduction group.
Preferably, said preparation comprises 0.05-2.0%w/v, more preferably the stabilizing agent of 0.08-1.0%w/v.
Particularly preferred stabilizing agent is Pluronic polyol, taurocholic acid and salt thereof and hydroxypropyl emthylcellulose.
The growth hormone liquid preparation of this stabilisation preferably also comprises the acceptable buffer of materia medica such as phosphate or citrate buffer, and its concentration is 2.5-50mM, more preferably is 10-20mM.
The pH of preparation is preferably 5.0-7.5, more preferably is 5.0-6.8, and more preferably 5.2-6.5 most preferably is 5.4-5.8.
In the growth hormone liquid preparation of preparation stabilisation, under growth hormone is not exposed to condition greater than the buffer concentration of the final buffer concentration in 2 * preparation, growth hormone is mixed with buffer, and follow under corresponding conditions, in mixture, add stabilizing agent.
In the preparation method of the growth hormone liquid preparation of especially preferred stabilisation, with the exposure limits of growth hormone concentration at 2 times the phosphate that is not more than each composition ultimate density or citrate buffer and Pluronic polyol.
The invention still further relates to a kind of treatment needs the human or animal patient's of growth hormone method, this method to comprise the as above stable growth hormone liquid preparation of general introduction of described patient's drug administration being learned effective dose.
Can carry out the injection of agglomerate or use the GH liquid preparation by using aerosol device or needleless injector by continuous intravenous infusion.
In this article, related " growth hormone " comprises the GH of all kinds, comprises people, cattle, pig, sheep and salmon and other, the biologically active derivatives of especially hGH, and GH.The GH derivant refers to the GH of the humans and animals kind that aminoacid sequence changes, as little aminoacid deletion or by other radical amino acid replacement.Also comprise the clipped form of GH and its derivant, and added amino acid whose GH, as methionyl hGH at proteic amino or carboxyl terminal.The hGH of another kind of type is modified to by forming (Davis etc., United States Patent (USP) 4,179,337) to active hGH aminoacid covalency addition Polyethylene Glycol.Detailed Description Of The Invention
The preparation method of the liquid preparation of GH provided by the invention and stabilizing agent produce a kind of on the solidification temperature and under be suitable for the storage of extended period and be suitable for treating the stable liquid GH preparation of using.The treatment preparation that comprises this stabilizing agent is stable, still allows treatment to use said preparation simultaneously.
According to a preferred embodiment of the present invention, GH is hGH.(1) human growth hormone's compositions
Term " human growth hormone " or " hGH " typical example be as by from natural origin, or the human growth hormone who produces by reconstitution cell culture system.The sequence of hGH and its feature description be at for example " hormonal medicaments ", Gueriguigan etc., and the USP meeting, Rockville is among the MD (1982).As mentioned above, this term also covers the biological activity equivalent that the different human growth hormone of one or more aminoacid is arranged in the hGH whole sequence, especially comprises met-hGH.This term also comprises amino acid whose variant of replacement, disappearance and insertion or the post translational modification thing of hGH.The hGH that is used for preparation of the present invention prepares by recombination method previously discussed usually.
GH especially " the materia medica effective dose " of hGH refers to produce the amount of therapeutic effect in various dosage regimens.Compositions of the present invention can be prepared to and comprise at least about 0.1mg/ml to the about 20mg/ml or the GH of volume more, is preferably extremely about 10mg/ml of about 1mg/ml, especially is preferably extremely about 5mg/ml of about 1mg/ml.(2) buffer and pH
Buffer can be the acceptable buffer agent of any materia medica, as phosphate, tris-HCl, citrate etc.Preferred buffer is phosphate or citrate buffer.Preferred buffer concentration less than 50mM, the most advantageously is 10-20mM more than or equal to 2mM.The suitable pH scope for preparing said preparation with buffer being used to thus of regulating is that about 5-is about 7.5, the most advantageously is about 5.6.Preparation pH should be less than 7.5 to reduce the desamidation of GH.(3) stabilizing agent
According to the present invention, said preparation comprises one or more stabilizing agents that is used to increase GH stability.This stabilizing agent can be polyethylene glycol-propylene glycol non-ionic surface active agent; as the Pluronic polyol; as Pluronics F127; F68, L64, PE6800 and PE6400; cholate; as the taurocholate or derivatives thereof, or the methylcellulose derivant, as hydroxypropyl emthylcellulose (HPMC).Said preparation can comprise two or more mixture of single stable agent or its.
The concentration of the stabilizing agent that adds will determine by selecting buffer and pH, but advantageously in the scope of 0.01%-5.0% (w/v), 0.05-2.0% more preferably, even more preferably 0.08-1.0%.When in a temperature range, comprise when freezing point experiences the storage period of prolongation up and down, or when said preparation stands interfacial stress, use stabilizing agent can increase stability of formulation.
Along with concentration increases, stabilizing agent increases the stability of preparation to interfacial stress.Yet the stabilizer concentration of increase reduces chemical stability.According to the present invention, with the stabilizer concentration optimization to obtain the high stability to interfacial stress, the chemical instability minimum of Zeng Jiaing simultaneously.(4) preferred formulation of stabilizing agent and hGH
In the preparation of preparation, in the hGH liquid preparation, add one or more stabilizing agents according to the present invention.As mentioned above, in process for preparation, growth hormone is exposed in the buffer concentration that is not more than 2 times of buffer ultimate densities, preferably adds stabilizing agent a moment before regulating final volume in preparation.The preparation that produces has the stability of increase to degeneration, and insensitive to undesirable reaction that may run in processing and storage.Adopt as this paper, term " processing " comprises filtration, packs in the bottle hGH solution and other operation that relates in producing preparation.
Can by hGH and stabilizing agent with required purity are mixed with physiologically acceptable excipient, buffer or antiseptic the liquid preparation for preparing the hGH that is used for the treatment of administration (the Remington pharmaceutical science, the 16th version, Osol, A. compiles (1980).Acceptable excipient be under concentration that adopts and dosage to nontoxic those of patient, comprise buffer agent, antiseptic, antioxidant, pH and osmotic pressure regulator.
If desired, the liquid preparation of growth hormone also can comprise one or more other stabilisation excipient.Other stabilisation excipient can comprise for example aminoacid, as glycine or alanine, and mannitol or other sugar alcohol or glycerol.In addition, liquid preparation can comprise other somatomedin, as insulin like growth factor or epidermal growth factor.
Preferred embodiment of the present invention provides a kind of method of effectively stablizing hGH.This preferred formulation comprises one or more stabilizing agents that is selected from Pluronic polyol, taurocholic acid or its salt or derivatives thereof and methylcellulose derivant.Preparation preferably comprises pure basically hGH, is not contained in pollution peptide or albumen or infectious substance that philtrum is found.The preparation of this preferred embodiment can comprise the materia medica acceptable additive in addition.This for example comprise buffer, etc. blend pH regulator agent, chelating agen, antiseptic, antioxidant, cosolvent etc., instantiation can comprise citrate, phosphate etc.When the expection application of preparation may damage aseptic, antiseptic can be added, in this case, the acceptable antiseptic of materia medica can be adopted, as benzyl alcohol or phenol.
Feasible can being extensive use of of the stability of the increase of the hGH that preparation prepared in accordance with the present invention provides may be than normally used those denseer hGH preparations when not having stabilizing agent.For example, the hGH liquid preparation of stabilisation also is reduced in the generation of degeneration of the spatial induction of the hGH that takes place in aerosolization or the needleless injection hGH preparation.Can carry out the further optimal allocation of hGH preparation, hGH preparation wherein of the present invention is by with the 1-50mg/ bottle, preferred 2-25mg/ bottle, and more preferably the amount of 3-10mg/ bottle is dispensed in the bottle.The stability that the hGH preparation increases makes can (most preferably be-20 ℃) under preference temperature such as freezing point, or on the freezing point preferred 2-8 ℃, be able to Long-term Storage when most preferably being 4 ℃.
The hGH preparation that is used for vivo medicine-feeding must be aseptic.By can easily reaching this point with the aseptic filtration membrane filtration.
Treatment is placed in the container with sterile access port usually with the hGH liquid preparation, for example intravenous solution bag or have the bottle of the stopper that can be passed by hypodermic needle.
Medicine-feeding way according to hGH liquid preparation of the present invention is consistent with known practice, for example by in the intravenous, intraperitoneal, brain, intramuscular, ophthalmic, intraarticular or intralesional injection or perfusion, or by continuous venoclysis.
Further aspect of the present invention will be conspicuous from the following examples and accompanying drawing.
Brief Description Of Drawings
Fig. 1 shows the chemical stability of hGH (1.5mg/ml) in 5mM phosphate buffer (pH6.0-7.5).
Fig. 2 shows that gathering by the inductive hGH of interfacial stress (vortex stirring) (at the 10mM acetate buffer, among the pH4.1-4.5, or at the 5mM phosphate buffer, the 2mg/ml solution among the pH6.0-7.5) is to the dependency of pH value of solution.
Fig. 3 illustrates the sedimentary ability of various stabilizing agents reduction interfacial stress (vortex stirring) inductive gathering hGH of having summed up.
Fig. 4 shows that difference only is that hGH imported two hGH (5mg/ml) preparations of the method in the excipient and has wherein added the 3rd the hGH stability of formulation of 0.005%w/v EDTA.
The method of EXAMPLE Example 1. screening stabilizing agents
Stabilizing agent reduce or prevention since the GH that interfacial stress causes especially the accumulative ability of hGH used quick method for congregating to obtain estimating, and analyze by size exclusion chromatography (SEC).Use placed in-line two TSK G3000SW posts (7.8mm internal diameter * 300mm, ToyoSodo, Japan) to carry out the chromatography of hGH.Mobile phase is made up of 0.1M phosphate (pH7.0) buffer, and pumps into 0.9ml/ minute flow velocity, by the eluent at the UV of 214nm absorbance detection hGH, uses the sample volume of 20 μ 1.
Fast method for congregating comprise by the polypropylene test tube of adding a cover (11mm in through * import high air/water termination second with constant speed vortex stirring hGH solution 15-60 in 74mm), under the room temperature, with sample balance 30 minutes so that precipitation take place, then, analyze filtrate with SEC with the little centrifugal filter paper filtering of 0.2 μ m cellulose acetate.The contrast solution of each sample of handling is not included in the SEC analysis.
The amount of remaining whole solubility hGH (peak area of monomer and high molecular weight material) is represented with the percent of the whole peak area (for hGH) of suitable untreated control solution.
Table 1 shows that various stabilizing agents are in the influence of pH7.0 by the inductive hGH aggregation extent of interfacial stress.
Table 2 shows various stable in the influence of pH6.0 by the inductive hGH aggregation extent of interfacial stress.
Table 3 shows that various stabilizing agents are in the influence of pH5.6 by the inductive hGH aggregation extent of interfacial stress.
Table 4 shows etc. and to ooze adjusting to the influence at hGH (1.5mg/ml) aggregation extent of pH5.6 in various buffer.
Table 5 shows that various stabilizing agents are in the influence of pH5.6 by freeze-thaw inductive hGH aggregation extent.
Shown in appended form, some excipient are to reducing or preventing because the inductive hGH of interfacial stress assembles very effective.It almost is quantitative protection that the Pluronic polyol provides when concentration is higher than 0.05%w/v, only residual monomer hGH.When taurocholic acid was higher than 0.02%w/v in concentration, providing almost was quantitative protection, only residual monomer hGH.When pH5.6, the deoxidation taurocholate is not suitable for as stabilizing agent, because it causes the hGH dimerization when not having interfacial stress.
Table 1 is at pH7.0 and when having excipient, by the gathering of the inductive hGH of surface stress (1.5mg/ml)
Table 2 is at pH6.0 and when having excipient, by the gathering of the inductive hGH of interfacial stress (1.5mg/ml)
Table 3 is at pH5.6 and when having excipient, by the gathering of the inductive hGH of interfacial stress (1.5mg/ml)
* there is the deoxidation taurocholate, the dimerization effect that takes place when not having interfacial stress
| Excipient | Concentration (mM) (%w/v) | The remaining all solubility hGH of % (n=2.SEC analysis) | The kind of remaining hGH |
| Buffer | Contrast | ????7.0±1.15 | (meansigma methods ± sd, n=5) |
| Pluronic polyol Pluronic F-127 | ????1.4??????1.75 ????3.2??????4.0 | ????100,99.8 ????100,100 | In all samples, only there is monomer |
| Taurocholic acid derivant: taurocholate deoxidation taurocholate | ????1.3??????0.07 ????5????????0.27 ????75???????4.0 ????0.4??????0.02 ????2????????0.11 ????30???????1.6 | ????75.3,68.5 ????100,100 ????100,100 ????13.7,15.0 ????100,100 ????100,100 | In all samples, only exist monomer in all samples, only to have monomer |
| Methylcellulose derivant: hydroxypropyl emthylcellulose (HPMC) | ????0.01?????0.1 ????0.05?????0.5 | ????44.1,47.1 ????98.9,99.4 | In all samples, only there is monomer |
| Excipient | Concentration (mM) (%w/v) | Remaining all solubility hGH of % (n=2, SEC analyzes) | Remaining hGH kind |
| Buffer | Contrast | 2.4±2.05 | (meansigma methods ± sd, n=6) |
| Pluronic polyol Pluronic F-127 Pluronic F-68 Pluronic L-64 Pluronic PE-6800 Pluronic PE-6400 | ?0.01????0.01 ?0.04????0.05 ?0.08????0.1 ?0.4?????0.5 ?1.6?????2.0 ?0.01????0.01 ?0.06????0.05 ?0.12????0.1 ?0.6?????0.5 ?0.03????0.01 ?0.17????0.05 ?0.35????0.1 ?1.7?????0.5 ?0.12????0.1 ?0.6?????0.5 ?2.4?????2.0 ?0.35????0.1 ?1.7?????0.5 | ?3.34,1.12 ?93.7,91.9 ?100,100 ?100,99.6 ?98.5,97.7 ?1.2,2.1 ?86.4,97.5 ?98.6,100 ?100,99.7 ?1.1,1.4 ?93.7,79.4 ?100,100 ?100,98.4 ?98.7,99.2 ?99.6,99.6 ?100,99.0 ?100,100 ?100,100 | In all samples, only exist monomer in all samples, only to exist monomer in all samples, only to exist monomer in all samples, only to exist monomer in all samples, only to have monomer |
| Taurocholic acid derivant: taurocholate deoxidation taurocholate | 1.3??????0.07 ?6???????0.34 ?15??????0.84 ?25??????1.4 ?0.4?????0.02 ?2???????0.10 ?6???????0.31 ?12??????0.63 | ?16.8,11.9 ?98.5,100 ?97.8,99.3 ?98.7,99.1 ?9.2,4.9 ?22.6,22.0 ?87.2,88.3 ?99.5,100 | In all samples, only there is about 4% dimer of about 0.5% dimer of monomer |
| Methylcellulose derivant: HPMC | ?0.01????0.1 ?0.03????0.25 ?0.04????0.4 | ?28.6,24.0 ?58.6,63.4 ?91.9,93.6 | In all samples, only there is monomer |
| Excipient | Concentration (mM) (%w/v) | Remaining all solubility hGH of % (n=2, SEC analyses) | The kind of remaining hGH |
| Buffer | Contrast | ?0.79,1.16 | (meansigma methods ± sd, n=2) |
| Pluronic polyhydric alcohol: Pluronic F-127 Pluronic F-68 | ?0.04???0.05 ?0.08???0.1 ?0.4????0.5 ?0.06???0.05 ?0.12???0.01 ?0.6????0.5 | ?69.6,53.8 ?99.5,99.7 ?99.5,98.8 ?69.9,66.8 ?99.3,98.1 ?99.8,100 | Exist 2% dimer, 0.7% dimer only to exist monomer in all samples, only to have monomer |
| Taurocholic acid derivant: taurocholate deoxidation taurocholic acid | ?5??????0.27 ?10?????0.54 ?25?????1.4 ?2??????0.10 ?6??????0.32 ?12?????0.63 | ?99.7,99.2 ?100,100 ?99.4,100 ?95.7,91.8 *?92.1,87.8 *?91.6,84.4 * | In all samples, only exist monomer to exist 7% dimer to exist 27% dimer to have 34% dimer |
| Methylcellulose derivant: HPMC | ?0.01????0.1 ?0.03????0.25 ?0.04????0.4 | ?5.8,2.1 ?34.8,36.7 ?88.8,88.4 | In all samples, only there is monomer |
Because aggregation is in the news and depends on phosphate concn (Pearlnan and Nguyen, 1992, J.Pharm.Pharmacol.44:178-185), studied add or do not add sodium chloride (to etc. ooze) citrate or phosphate (5 or 20mM) buffer in hGH (1.5mg/ml, aggregation properties pH5.6).
According to foregoing experimental technique, only change the processing time (15 seconds).Table 4 is in the gathering of the hGH (1.5mg/ml) of pH5.6 in various buffer solution systems
The a monomer adds high molecular weight species b hGH dissolubility deficiency.
| Buffer solution system | Remaining all the solubility hGH of % do not add NaCl and add NaCl | The kind of the hGH that exists | |
| 5 mM phosphate | _b | ?44.5,30.3 | Only there is |
| 5 mM citrates | 18.9,21.2 | ?44.3,46.4 | Only there is monomer |
| 20 mM phosphate | 28.5,33.1 | ?43.1,38.7 | Only there is monomer |
| The 20mM citrate | 24.6,33.2 | ?55.5,50.0 | Only there is monomer |
Do not find that hGH assembles character or the buffer concentration that depends on buffer.HGH assembles and ionic strength (when regulating with NaCl) negative correlation.
Studied when having excipient, assembled by freeze-thaw inductive hGH (ooze be 1.5mg/ml among the citrate buffer pH5.6 at 20mM etc.).In the time of-20 ℃, when having various excipient,, then melt under the room temperature, and balance 30 minutes is to allow precipitation produce freezing 24 hours of hGH sample (100 μ l).Carry out the analysis of filtered sample as previously mentioned by SEC.
Table 5 is at pH5.6 and when having excipient, by the gathering of freeze-thaw inductive hGH (1.5mg/ml)
| Excipient | Concentration (mM) (%w/v) | Remaining all solubility hGH of % (n=2, SEC analyzes) | The kind of remaining hGH |
| Buffer | Contrast | 98.8,95.5 | Only there is monomer |
| Pluronic polyol Pluronic F-127 Pluronic F-68 | ?0.08?????0.1 ?0.12?????0.1 | ?95.9,97.5 ?100,100 | Only exist monomer only to have monomer |
| Taurocholic acid derivant: taurocholate deoxidation taurocholate | ?5????????0.27 ?2????????0.1 | ?97.9,100 ?95.7,91.8 | Only exist monomer to comprise 24% dimer |
| Methylcellulose derivant: HPMC | ?0.03?????0.25 | ?97.2,97.3 | Only there is monomer |
HGH after freeze-thaw assembles not extensive, but does not increase with the adding of excipient.Embodiment 2
Fig. 1 is the representative scattergram of hGH (15mg/ml) in the chemical stability of 5mM phosphate buffer pH6.0-7.5 (being housed in 40 ℃).According to the method for describing in the American Pharmacopeia (USP1990), use Vydac C-4 post to analyze degradation samples by reversed-phase high-performance liquid chromatography (RP-HPLC).Determine that according to the method for describing in the American Pharmacopeia advance notice (nineteen ninety 11-12 day) catabolite is deacylated tRNA amine hGH or oxidation hGH.Being present in natural hGH (figure A), deacylated tRNA amine hGH (figure B) and the amount of oxidation hGH (scheme C) of degraded in the sample uses with respect to (natural hGH or degradation product) peak area percent of the whole peak area (for hGH) of pH6.0 (zero), pH6.5 (●), pH7.0 () and pH7.5 () and represents.
Find to follow first order kinetics in the pH scope that is lost in 6.0-7.5 of natural hGH, in 8-40 ℃ temperature range, follow the Arrhenius behavior.Find 40 ℃ first order rate constant when pH6.0 2.4 * 10
-2My god
-1During to pH7.5 7.4 * 10
-2My god
-1Scope in.Consistent with disclosed report (Pearlman and Nguyen, 1989, see above), desamidation and Oxidation are the main paties of hGH degraded.Deacylated tRNA amine hGH forms with the speed that forms greater than oxidation hGH, pH value be 6.0 or when lower chemical stability strengthen.
Fig. 2 shows and uses as the method for description among the embodiment 1, by interfacial stress inductive in 10mM acetate buffer (pH4.1-4.5) or 5mM phosphate buffer (pH6.6-7.5) gathering and the precipitation of hGH (2mg/ml).The amount of monomer hGH (because monomeric peak area) or remaining whole solubility hGH (because monomer adds the peak area of more high-grade aggregation) is used with respect to the percent of the peak area of suitable untreated control solution and is represented.Data are illustrated in the amount that vortex stirs the remaining soluble and monomeric hGH in 30 seconds (zero) or 60 seconds (●) back (figure A) or whole solubility hGH (figure B).
In the zone of pH5-6, the aggregation of hGH and precipitation maximum afterwards.In the pH4.1-6.0 scope, after interfacial stress, only monomer hGH is retained in the solution.In the pH7.0-7.5 scope, mainly exist solubility to assemble thing (dimer and more high grade collecting thing).
Fig. 3 shows that excipient (%w/v) stirs the influence by the aggregation of the inductive hGH of interfacial stress (being 1.5mg/ml) in 60 seconds to passing through as described in Figure 2 with the constant speed vortex in 5mM phosphate buffer pH5.6.Data are illustrated in when having Pluronic F-68 (zero), Pluronic F-127 (●), sodium taurocholate () or HPMC (), the percent of the whole solubility hGH (monomer adds more high-grade aggregation) that still keep after the processing of representing with the percent of the hGH peak area analyzed with respect to SEC in the suitable contrast solution.
When not having excipient, the hGH less than 1% still is retained in the solution.Adding Pluronic polyol, taurocholate or HPMC causes remaining solubility hGH to roll up.Especially to provide almost be quantitative anti-gathering protection for Pluronics F-68 and F-127 and taurocholic acid salt pair hGH.Embodiment 3
Fig. 4 shows the influence of compound method to the hGH preparation stability.By with the hGH solution concentration of purification to 7-7.5mg/ml, and add comprising all excipient and being adjusted to the solution of certain pH of 2 times of concentration, described pH makes need not further regulate the liquid preparation that can produce pH5.6, and last water is regulated with the final hGH concentration that obtains 5mg/ml and come formulated 1.
By buffer-exchanged, by in the buffer that exchanges to all excipient (except Pluronic F-68) that comprise desired concn hGH solution concentration to the desired concn of purification being come formulated 2.The solid Pluronic F-68 that then adds capacity is to produce desired concn.Check pH subsequently, regulate pH if desired.
HGH (growth hormone) 5mg
Citric acid-hydrate 2.04mg/ml
Citrate trisodium dihydrate 2.85mg/ml
Sodium chloride 6.23mg/ml
Sodium hydroxide 0.388mg/ml
Benzyl alcohol 0.9%
Pluronic?F-68????????0.08％
pH???????????????????5.6
Formulated 3 as preparation 2, and it comprises and preparation 1 and 2 identical compositions, and is added with 0.005%w/vEDTA.
Under 40 ℃, preserve preparation 1-3, and analyze hGH content by size exclusion HPLC with 40 days interval.In this accelerated stability test of 40 ℃, especially compare with preparation 1 as shown in Figure 4, preparation 2 and 3 demonstrates better stability.
Claims (11)
1. method for preparing stable growth hormone liquid preparation; what wherein said preparation comprised growth hormone, buffer and stabilisation effective dose at least aly is selected from following stabilizing agent: (ⅰ) polyethylene glycol-propylene glycol non-ionic surface active agent; (ⅱ) taurocholic acid or its salt or derivant; (ⅲ) methylcellulose derivant; wherein this method is included in growth hormone and is not exposed under the condition greater than the buffer of buffer in 2 * preparation or stabilizing agent ultimate density or stabilizer concentration, and growth hormone is mixed with buffer and stabilizing agent.
2. according to the process of claim 1 wherein that growth hormone is the human growth hormone.
3. according to the method for claim 1 or 2, wherein said preparation comprises 0.1-5.0%w/v, preferred 0.05-2.0%w/v, the more preferably stabilizing agent of 0.08-1.0%w/v.
4. according to each method of claim 1-3, wherein stabilizing agent is selected from Pluronic polyol, taurocholic acid and its salt, and hydroxypropyl emthylcellulose.
5. according to each method of claim 1-4, wherein preparation comprises the Pluronic polyol of 0.08%w/v as stabilizing agent.
6. according to each method of claim 1-5, wherein said preparation comprises the acceptable buffer of materia medica, preferably phosphate or citrate buffer, and its concentration is 2.5-5.0mM, is preferably 10-20mM.
7. according to each method of claim 1-6, wherein the pH of preparation is 5.0-7.5, is preferably 5.0-6.8, and more preferably 5.2-6.5 most preferably is 5.4-5.8.
8. according to each method of claim 1-7, wherein stabilizing agent is added in the preparation a moment before regulating final volume.
9. stable growth hormone liquid preparation, it is by according to each method preparation of claim 1-8.
10. stable growth hormone liquid preparation, it comprises growth hormone, buffer and stabilisation effective dose at least a taurocholic acid or its salt or derivant, and the stabilizing agent of methylcellulose derivant of being selected from.
11. a treatment needs the human or animal patient's of growth hormone method, it comprises the preparation according to claim 9 or claim 10 of described patient's drug administration being learned effective dose.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AUPN8012 | 1996-02-12 | ||
| AUPN8012A AUPN801296A0 (en) | 1996-02-12 | 1996-02-12 | Stabilised growth hormone formulation and method of preparation thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1214633A true CN1214633A (en) | 1999-04-21 |
| CN1132626C CN1132626C (en) | 2003-12-31 |
Family
ID=3792327
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN97193316A Expired - Fee Related CN1132626C (en) | 1996-02-12 | 1997-02-12 | Stabilised growth hormone formulation and method of preparation thereof |
Country Status (13)
| Country | Link |
|---|---|
| US (1) | US6593296B1 (en) |
| EP (1) | EP0889733B1 (en) |
| JP (1) | JP4255515B2 (en) |
| CN (1) | CN1132626C (en) |
| AT (1) | ATE340587T1 (en) |
| AU (1) | AUPN801296A0 (en) |
| CA (1) | CA2246501C (en) |
| DE (1) | DE69736739T2 (en) |
| DK (1) | DK0889733T3 (en) |
| HK (1) | HK1017851A1 (en) |
| RU (1) | RU2191029C2 (en) |
| TR (1) | TR199801558T2 (en) |
| WO (1) | WO1997029767A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659701A (en) * | 2002-12-31 | 2010-03-03 | 阿尔特斯制药公司 | Human growth hormone crystals and methods for preparing them |
| CN105363022A (en) * | 2006-07-06 | 2016-03-02 | 株式会社大熊 | Stable human growth hormone liquid preparation |
| CN105925545A (en) * | 2008-04-21 | 2016-09-07 | 诺沃—诺迪斯克保健股份有限公司 | Dry Transglutaminase Composition |
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| US6277410B1 (en) * | 1992-10-08 | 2001-08-21 | Supratek Pharma Inc. | Copolymer compositions for oral delivery |
| JP2003504346A (en) * | 1999-07-12 | 2003-02-04 | グランデイス・バイオテツク・ゲー・エム・ベー・ハー | Growth hormone preparations |
| AT408721B (en) * | 1999-10-01 | 2002-02-25 | Cistem Biotechnologies Gmbh | PHARMACEUTICAL COMPOSITION CONTAINING AN ANTIG |
| KR100890679B1 (en) | 2000-02-24 | 2009-03-26 | 일라이 릴리 앤드 캄파니 | Non-aqueous injectable formulations for extended release of somatotropin |
| US6719992B2 (en) | 2000-06-26 | 2004-04-13 | Monsanto Technology Llc | Non-aqueous surfactant-containing formulations for extended release of somatotropin |
| US6664234B1 (en) | 2000-06-30 | 2003-12-16 | Monsanto Technology Llc | Non-aqueous injectable formulation preparation with pH adjusted for extended release of somatotropin |
| GB2371227A (en) * | 2001-01-10 | 2002-07-24 | Grandis Biotech Gmbh | Crystallisation - resistant aqueous growth hormone formulations |
| JP4610154B2 (en) * | 2002-05-30 | 2011-01-12 | 大塚製薬株式会社 | Injectable preparation |
| US20060252682A1 (en) * | 2003-03-18 | 2006-11-09 | Ares Trading S.A. | Liquid growth hormone formulation and process of preparation thereof |
| CA2540172A1 (en) * | 2003-09-25 | 2005-03-31 | Cangene Corporation | Liquid human growth hormone formulation containing polyethylene glycol |
| JP4845741B2 (en) * | 2003-12-23 | 2011-12-28 | ファルマシア コーポレーション | Stable growth hormone liquid formulation |
| ES2383994T3 (en) * | 2004-04-07 | 2012-06-28 | Ares Trading S.A. | Liquid growth hormone formulation |
| KR101173717B1 (en) | 2004-11-09 | 2012-08-13 | 아레스 트레이딩 에스.에이. | Method for purifying fsh |
| JP4944112B2 (en) | 2005-08-26 | 2012-05-30 | アレス トレーディング ソシエテ アノニム | Process for the preparation of glycosylated interferon beta |
| EP1960419B1 (en) | 2005-12-09 | 2016-03-16 | Ares Trading S.A. | Method for purifying fsh or a fsh mutant |
| EP2049148B1 (en) * | 2006-07-06 | 2016-09-28 | Daewoong Co., Ltd. | A stable liquid formulation of human growth hormone |
| WO2013014196A1 (en) * | 2011-07-25 | 2013-01-31 | Sandoz Ag | Aqueous formulation comprising at least a neutral salt and a biopharmaceutical protein |
| US10064951B2 (en) * | 2012-03-30 | 2018-09-04 | Hanmi Science Co., Ltd. | Liquid formulation of highly concentrated long-acting human growth hormone conjugate |
| CA2925416A1 (en) * | 2013-09-27 | 2015-04-02 | Hanmi Pharm. Co., Ltd. | Sustained type human growth hormone preparation |
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| AU6073386A (en) | 1985-07-30 | 1987-02-05 | International Minerals & Chemical Corporation | Stabilization of growth promoting hormones |
| IT1222395B (en) | 1987-07-30 | 1990-09-05 | Pierrel Spa | PHARMACEUTICAL COMPOSITION FOR INTRANASAL ADMINISTRATION INCLUDING THE HORMONE GHRH, A COLINERGIC AGONIST AND / OR A BILE SALT |
| DE3782737T3 (en) | 1987-08-21 | 1999-05-20 | Imcera Group Inc., Northbrook, Ill. | Stabilization of growth hormones. |
| US5096885A (en) * | 1988-04-15 | 1992-03-17 | Genentech, Inc. | Human growth hormone formulation |
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| EP0374120A3 (en) * | 1988-12-13 | 1991-07-31 | Monsanto Company | Comosition for controlled release of polypeptides |
| US5013714A (en) * | 1988-12-15 | 1991-05-07 | Lindstrom Richard L | Viscoelastic solution |
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| US5182258A (en) * | 1989-03-20 | 1993-01-26 | Orbon Corporation | Systemic delivery of polypeptides through the eye |
| US5126324A (en) * | 1990-06-07 | 1992-06-30 | Genentech, Inc. | Method of enhancing growth in patients using combination therapy |
| US5210017A (en) * | 1990-11-19 | 1993-05-11 | Genentech, Inc. | Ligand-mediated immunofunctional hormone binding protein assay method |
| SE9201073D0 (en) | 1992-04-03 | 1992-04-03 | Kabi Pharmacia Ab | PROTEIN FORMULATION |
| CA2489978A1 (en) * | 1992-07-31 | 1994-02-17 | Genentech, Inc. | Human growth hormone aqueous formulation |
| EP1652534A3 (en) * | 1992-10-02 | 2007-05-02 | Genetics Institute, LLC | Composition comprising coagulation factor VIII formulation, process for its preparation and use of a surfactant as stabilizer |
| NZ262634A (en) * | 1993-02-23 | 1997-02-24 | Genentech Inc | Stabilizing polypeptides against degradation by organic solvents by admixing the peptide with trehalose or mannitol |
| EP0736041B1 (en) * | 1993-11-17 | 2006-02-08 | Athena Neurosciences, Inc. | Transparent liquid for encapsulated drug delivery |
-
1996
- 1996-02-12 AU AUPN8012A patent/AUPN801296A0/en not_active Abandoned
-
1997
- 1997-02-12 DE DE69736739T patent/DE69736739T2/en not_active Expired - Lifetime
- 1997-02-12 CA CA2246501A patent/CA2246501C/en not_active Expired - Fee Related
- 1997-02-12 WO PCT/AU1997/000075 patent/WO1997029767A1/en active IP Right Grant
- 1997-02-12 AT AT97902109T patent/ATE340587T1/en active
- 1997-02-12 DK DK97902109T patent/DK0889733T3/en active
- 1997-02-12 RU RU98117135/14A patent/RU2191029C2/en active
- 1997-02-12 TR TR1998/01558T patent/TR199801558T2/en unknown
- 1997-02-12 EP EP97902109A patent/EP0889733B1/en not_active Revoked
- 1997-02-12 US US09/125,267 patent/US6593296B1/en not_active Expired - Fee Related
- 1997-02-12 JP JP52882197A patent/JP4255515B2/en not_active Expired - Lifetime
- 1997-02-12 CN CN97193316A patent/CN1132626C/en not_active Expired - Fee Related
-
1999
- 1999-07-09 HK HK99102933A patent/HK1017851A1/en not_active IP Right Cessation
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659701A (en) * | 2002-12-31 | 2010-03-03 | 阿尔特斯制药公司 | Human growth hormone crystals and methods for preparing them |
| CN101659701B (en) * | 2002-12-31 | 2019-03-26 | 阿尔特斯制药公司 | Human growth hormone crystals and the method for preparing them |
| CN105363022A (en) * | 2006-07-06 | 2016-03-02 | 株式会社大熊 | Stable human growth hormone liquid preparation |
| CN105925545A (en) * | 2008-04-21 | 2016-09-07 | 诺沃—诺迪斯克保健股份有限公司 | Dry Transglutaminase Composition |
| US10391062B2 (en) | 2008-04-21 | 2019-08-27 | Novo Nordisk Healthcare Ag | Dry transglutaminase composition |
Also Published As
| Publication number | Publication date |
|---|---|
| RU2191029C2 (en) | 2002-10-20 |
| AUPN801296A0 (en) | 1996-03-07 |
| EP0889733B1 (en) | 2006-09-27 |
| DE69736739D1 (en) | 2006-11-09 |
| ATE340587T1 (en) | 2006-10-15 |
| WO1997029767A1 (en) | 1997-08-21 |
| EP0889733A4 (en) | 1999-11-03 |
| JP4255515B2 (en) | 2009-04-15 |
| EP0889733A1 (en) | 1999-01-13 |
| DE69736739T2 (en) | 2007-08-16 |
| TR199801558T2 (en) | 1998-11-23 |
| US6593296B1 (en) | 2003-07-15 |
| CA2246501C (en) | 2012-07-10 |
| HK1017851A1 (en) | 1999-12-03 |
| CN1132626C (en) | 2003-12-31 |
| CA2246501A1 (en) | 1997-08-21 |
| JP2000504696A (en) | 2000-04-18 |
| DK0889733T3 (en) | 2007-01-29 |
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